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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Prokaryotic Expression and Antigenic Characterization of Three Recombinant Leishmania Antigens for Serological Diagnosis of Canine Leishmaniasis
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Prokaryotic Expression and Antigenic Characterization of Three Recombinant Leishmania Antigens for Serological Diagnosis of Canine Leishmaniasis

机译：三种重组利什曼原虫抗原的原核表达和抗原学特性对犬利什曼病的血清学诊断

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Three recombinant antigens of Leishmania chagasi (= L. infantum) were expressed in prokaryotic systems and evaluated (using a panel of dog sera characterized by parasitological and serological immunofluorescent antibody test [IFAT] techniques) as diagnostic markers of infection. The whole open reading frame encoding K9, the gene fragment encoding the repetitive sequence of K26, and the 3′-terminal gene fragment encoding a single 39-amino-acid subunit of the kinesin-related protein K39 (K39sub) were amplified from L. infantum DNA and cloned into a pGEX-2T expression vector in frame with glutathione S-transferase (GST). The sensitivity and specificity of enzyme-linked immunosorbent assays (ELISAs) using K26 as an antigen (evaluated with sera from 20 parasitologically positive and 20 parasitologically negative dogs) were both 100% (95% confidence interval [CI] = 83.2 to 100). When K9 and K39sub were used, sensitivity was 95% (95% CI = 75.1 to 99.9) and specificity was 100% (95% CI = 83.2 to 100). Using 182 field sera, a good agreement was found between the recombinant K26 ELISA and IFAT (K = 0.92; 95% CI = 0.86 to 0.98) results and between the K9 and K39sub ELISA (used in parallel) and IFAT (K = 0.87; 95% CI = 0.80 to 0.95) results. The results demonstrate that each antigen carries immunodominant epitopes and that their combination may further increase the sensitivity of currently available serological tests.
机译：在原核系统中表达了三种（= L。infantum ）重组抗原，并对其进行了评估（使用一组具有寄生虫学和血清学免疫荧光抗体试验[IFAT]的狗血清）技术）作为感染的诊断标记。从扩增了编码K9的完整开放阅读框，编码K26重复序列的基因片段以及编码驱动蛋白相关蛋白K39（K39sub）的单个39个氨基酸亚基的3'-末端基因片段。 > L。婴儿DNA ，并与谷胱甘肽 S -转移酶（GST）一起克隆入pGEX-2T表达载体。使用K26作为抗原的酶联免疫吸附测定（ELISA）的敏感性和特异性（用来自20株寄生虫阳性和20株寄生虫阴性的犬的血清进行评估）均为100％（95％置信区间[CI] = 83.2至100）。使用K9和K39sub时，敏感性为95％（95％CI = 75.1至99.9），特异性为100％（95％CI = 83.2至100）。使用182场血清，在重组K26 ELISA和IFAT（ K = 0.92; 95％CI = 0.86至0.98）和在K9和K39sub ELISA（并行使用）之间发现了很好的一致性和IFAT（ K = 0.87; 95％CI = 0.80至0.95）。结果表明，每种抗原均带有免疫优势表位，并且它们的组合可能会进一步提高当前可用血清学检测的敏感性。

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